The major objective of this project is the detection of Mycobacterium tuberculosis (Mtb) in biological samples (sputum, tissue samples, and other fluids). This will be accomplished by: 1. Cloning unique fragments of Mycobacterium tuberculosis in the vector pUC9. 2. Sequencing several unique Mtb DNA sequences for the development of target sequences for polymerase chain amplification. 3. Amplifying the test Mtb targets on purified DNA from the Mtb family of sequences, the M. avium-intracellulare group of mycobacteria and other closely related species. 4. Apply the Mtb targets to standard cultures of Mtb to test sensitivity. 5. Lastly, attempt Mtb detection with clinical specimens. Within six months, it is expected that target DNA sequences will be characterized with respect to specificity to mycobacterial strains.